Brains of wild-type C57BL/6N mice were dissected within 24 h after birth (postnatal days 0–1), and the cortices were isolated, transferred into HBSS-containing tubes, and treated with PDD solution (HBSS -/-, 0.01% papain (w/v) (Worthington Biochemical Corporation, Lakewood, NJ, USA), 0.1% (w/v) dispase II (Hoffmann-La Roche AG, Basel, Switzerland), 0.01% (w/v) DNase I (Worthington Biochemical Corporation), 12.4 mM MgSO4) [54 (link)], followed by mechanical trituration. A total of 2 × 105 cells per well were seeded in an NBA-media mix (Neurobasal A, 1% (v/v) GlutaMax, 2% (v/v) B27 50x, 1% (v/v) sodium pyruvate, 1% (v/v) antibiotic/antimycotic (all from Gibco/Thermo Scientific, Schwerte, Germany)) in 12-well plates coated with poly-L-lysine (2.5 mg/mL poly-L-lysine (Sigma-Aldrich, Munich, Germany) in 150 mM borate buffer pH 8.4). Cells were cultured for one week at 37 °C in a 5% CO2 humidified atmosphere and treated as indicated with 10 µM fluoxetine [15 (link)].
RNA isolation was performed with the Purelink RNA Kit from Thermo Scientific (Schwerte, Germany), according to the manufacturer’s protocol. RNA qualities and concentrations were assessed using a NanoDrop ND-1000 UV-Vis spectrophotometer (Peqlab, Erlangen, Germany). Five hundred nanograms of RNA were reverse transcribed into cDNA using the Quanta cDNA Kit (Gaithersburg, MD, USA), according to the manufacturer’s protocol.
RNA isolation was performed with the Purelink RNA Kit from Thermo Scientific (Schwerte, Germany), according to the manufacturer’s protocol. RNA qualities and concentrations were assessed using a NanoDrop ND-1000 UV-Vis spectrophotometer (Peqlab, Erlangen, Germany). Five hundred nanograms of RNA were reverse transcribed into cDNA using the Quanta cDNA Kit (Gaithersburg, MD, USA), according to the manufacturer’s protocol.
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